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. 2010 Nov 4;6(11):e1001196. doi: 10.1371/journal.pgen.1001196

Figure 3. DIM-5 interacts with CUL4, DDB1, and DIM-9.

Figure 3

(A) Southern hybridization with a probe corresponding to the normally methylated 8:A6 region was performed using genomic DNA from the indicated strains digested with the cytosine-methylation-sensitive BfuCI (B) and –insensitive DpnII (D) endonucleases. (B) Histones extracted from wildtype, dim-5, dim-7, dim-8, cul4 and dim-9 were subjected to western blotting using antibodies to H3K9me3 and H3K4me2 as indicated. (C) Immunoprecipitation experiments were performed using extracts from strains expressing FLAG-CUL4 or DDB1-FLAG (+). The input fraction, the α-FLAG immunoprecipitate (IP:αFLAG), the α-DIM-5 immunoprecipitate (IP: α-DIM-5) and the mock immunoprecipitate (IP: mock) were subject to western blotting and probed with the α-FLAG or α-DIM-5 antibodies as indicated (WB). The asterisk indicates a non-specific cross-reacting band. (D) Immunoprecipitation experiments were performed using extracts from strains expressing DIM-9-FLAG, DIM-5-HA, or both (indicated by + or −). The input fraction, the α-HA immunoprecipitate fraction (IP:α-HA) and the α-FLAG immunoprecipitate fraction (IP:α-FLAG) were subjected to western blotting and probed with the α-FLAG or α-HA antibodies as indicated (WB).